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Meta-analysis was used to compare yield protection and nematode suppression provided by two seed-applied and two soil-applied nematicides against Meloidogyne incognita and Rotylenchulus reniformis on cotton across 3 years and several trial locations in the U.S. Cotton Belt. Nematicides consisted of thiodicarb- and fluopyram-treated seed, aldicarb and fluopyram applied in furrow, and combinations of the seed treatments and soil-applied fluopyram. The nematicides had no effect on nematode reproduction or root infection but had a significant impact on seed cotton yield response ([Formula: see text]), with an average increase of 176 and 197 kg/ha relative to the nontreated control in M. incognita and R. reniformis infested fields, respectively. However, because of significant variation in yield protection and nematode suppression by nematicides, five or six moderator variables (cultivar resistance [M. incognita only], nematode infestation level, nematicide treatment, application method, trial location, and growing season) were used depending on nematode species. In M. incognita-infested fields, greater yield protection was observed with nematicides applied in furrow and with seed-applied + in-furrow than with solo seed-applied nematicide applications. Most notable of these in-furrow nematicides were aldicarb and fluopyram (>131 g/ha) with or without a seed-applied nematicide compared with thiodicarb. In R. reniformis-infested fields, moderator variables provided no further explanation of the variation in yield response produced by nematicides. Furthermore, moderator variables provided little explanation of the variation in nematode suppression by nematicides in M. incognita- and R. reniformis-infested fields. The limited explanation by the moderator variables on the field efficacy of nematicides in M. incognita- and R. reniformis-infested fields demonstrates the difficulty of managing these pathogens with nonfumigant nematicides across the U.S. Cotton Belt.

In this study we describe an improved cepstral method that can be used for estimating the depth (

From 2015 to 2020, 342 isolates of Xanthomonas citri pv. malvacearum (Xcm) were obtained from cotton fields in Texas, including 64 isolates collected from symptomatic cultivars that were thought to be resistant to race 18 strains of Xcm (the predominant race in the United States). Symptoms on highly resistant cultivars prompted concern that a new race of Xcm was present. The 342 isolates were inoculated on a race 18 susceptible (DP 1747NR B2XF) and race 18 resistant (S295) cotton cultivar, and none of the isolates caused blight-type symptoms (water soaking and chlorosis) on S295, indicating that the B12 resistant gene was still beneficial for disease management. Four cultivars, varying in their field response to bacterial blight, were inoculated with each of 17 isolates of Xcm, and the incidence of plants exhibiting bacterial blight symptoms averaged 87% for DP 1747NR B2XF, 51% for partially susceptible NG 4936 B3XF, 16% for partially resistant DP 1646 B2XF, and 0% for S295. Xcm isolates from Texas (11), Georgia (1), and Oklahoma (1) were sequenced, and their type three effectors (T3Es) were predicted. All isolates (Georgia, Oklahoma, and Texas) had the same T3E proteins as previously identified Xcm race 18 isolates (tested for 25 genes), including XopJ. Race 1, 2, 3, and 12 of Xcm included in the comparisons were all missing the XopJ gene. Use of cultivars with the B12 gene is an effective strategy to manage bacterial blight of cotton.

A greenhouse experiment was conducted to test the hypothesis that cultivar use contributed to historical shifts in disease predominance patterns in the southeastern United States over the past 50 years. Passalora arachidicola (Pa), the causal agent of early leaf spot (ELS), and Nothopassalora personata (Np), the causal agent of late leaf spot (LLS), were inoculated separately or together on three historically dominant cultivars, Florunner (1970 to 1996), Georgia Green (1996 to 2008) and Georgia-06G (2008 to present), and on one susceptible cultivar, Georgia Valencia. These results suggest that the transition from Florunner to Georgia Green may have contributed to the historical shift from LLS to ELS predominance observed in the 1990s, with sporulation potential as a possible mechanistic explanation. There was no evidence that Georgia-06G contributed to the resurgence of LLS in recent years. A negative association between ELS and LLS, where LLS is more suppressed in the presence of ELS, demonstrates that the dynamics of the ELS-LLS disease patterns is complex. Understanding factors that contribute to disease predominance will improve predictive abilities and support the development of cultural practices and fungicide programs specific to which pathogen is expected to dominate.

Cotton bacterial blight (CBB), caused by Xanthomonas citri pv. malvacearum, was a major disease of cotton in the United States in the early part of the twentieth century. The reemergence of CBB revealed many gaps in our understanding of this important disease. In this study, we employed a wild-type (WT) field isolate of X. citri pv. malvacearum from Georgia (U.S.A.) to generate a nonpathogenic hrcV mutant lacking a functional type-III secretion system (T3SS−). We tagged the WT and T3SS− strains with an auto-bioluminescent Tn7 reporter and compared colonization patterns of CBB-susceptible and CBB-resistant cotton seedlings using macroscopic image analysis and bacterial load enumeration. WT and T3SS− X. citri pv. malvacearum strains colonized cotton cotyledons of CBB-resistant and CBB-susceptible cotton cultivars. However, X. citri pv. malvacearum populations were significantly higher in CBB-susceptible seedlings inoculated with the WT strain. Additionally, WT and T3SS− X. citri pv. malvacearum strains systemically colonized true leaves, although at different rates. Finally, we observed that seed-to-seedling transmission of X. citri pv. malvacearum may involve systemic spread through the vascular tissue of cotton plants. These findings yield novel insights into potential X. citri pv. malvacearum reservoirs for CBB outbreaks.

Soybean fields in Georgia were surveyed for plant-parasitic nematodes (PPNs) during the 2021 growing season. The frequency and population density of ten PPN taxa were determined in 144 soil samples collected from soybean fields located within 31 Georgia counties. Between 1-6 PPN taxa were identified in 97% of the soil samples with >50% of samples harboring four or more taxa. Spiral nematode (Helicotylenchus spp.) was the most prevalent nematode (85%), followed by lesion (Pratylenchus spp., 71%), stubby root (Trichodoridae, 69%), ring (Criconematidae, 54%), root-knot (Meloidogyne spp., 49%), stunt (Tylenchorhynchus spp., 11%), reniform (Rotylenchulus spp., 8%), cyst (Heteroderidae, 8%), dagger (Xiphinema spp., 4%), and lance (Hopolaimus spp., 2%). Fourteen soil samples from nine Georgia counties were confirmed to have the soybean cyst nematode, Heterodera glycines (SCN), based on morphological identification, molecular identification, and evidence of reproduction on soybean. HG type tests determined the virulence of each SCN population on resistant soybean. All SCN populations tested were able to reproduce at a female index ≥10% on PI 88788. HG type 2.- was the most frequent (57%), followed by HG type 1.2.- (29%) and HG type 1.2.3.- (14%). Meloidogyne incognita was the most frequently identified species of root-knot nematode (80%), followed by M. arenaria (28%), and M. javanica (6%). Our survey results serve to 1) raise awareness of PPNs among Georgia soybean producers to inform future management decisions and 2) improve information available to soybean breeders as they move forward with their efforts to incorporate multi-nematode resistance into improved soybean cultivars.

Management of fungicide resistance in Nothopassalora personata, the cause of late leaf spot of peanut (Arachis hypogaea), is a challenge in peanut production areas of the southeastern United States. Field experiments were conducted in Tifton, GA in 2020 and 2021, and in Plains, GA in 2021 to determine the effects of mixtures of seven elemental sulfur fungicide products with the QoI fungicide azoxystrobin on late leaf spot in fields where QoI-resistant populations of N. personata are suspected. Application of azoxystrobin or sulfur alone had little reduction in on standardized area under the disease progress curve (SAUDPC) or final disease intensity ratings compared to the nontreated control. However, mixtures of azoxystrobin with all micronized sulfur products evaluated had SAUDPC values and final disease intensity ratings lower less than those in the azoxystrobin alone or Microthiol Disperss 80W sulfur alone. One non-micronized sulfur formulation had SAUDPC values and final intensity ratings similar to those of azoxystrobin or Microthiol Disperss 80W sulfur alone. The results suggest multiple micronized elemental sulfur products have potential for use as mixing partners with azoxystrobin leaf spot control in fields where the efficacy of azoxystrobin alone is not adequate.

Growers have rapidly adopted auxin-resistant cotton and soybean technologies. In Georgia, growers who plant auxin-resistant cotton/soybean are required to utilize nozzles that produce larger (coarser) droplets when spraying auxin herbicides to minimize potential off-target movement of pesticides. Consequently, these nozzles are also used in peanut (an important rotational crop with cotton) since changing nozzles between crops is uncommon for growers. However, larger droplets can result in reduced spray coverage which may lead to less effective pest control. Therefore, seven on-farm trials were conducted in commercial peanut fields using commercial sprayers from 2018 to 2020 across four different locations in Georgia to compare the spray performance of air-induction (AI) nozzles that produce very coarse to ultra coarse droplets (VMD50 ≥ 404 microns) with non-AI (conventional flat fan) nozzles that produce medium to coarse droplets (403≥VMD50≥236 microns) for pest management in peanuts. For each trial, test treatments were implemented in large replicated strips where each strip represented a nozzle type. For nozzle comparison, XR and XRC represented non-AI nozzles while TADF, TDXL, TTI, and TTI60 represented the commonly used AI nozzles in these trials. Spray deposition data for each nozzle along with disease ratings, weed and insect control ratings were collected in all on-farm trials. Peanut yield was collected at harvest. Results indicated that the AI nozzles produced larger droplets than the non-AI nozzles in all nozzle tests; however, the spray coverage varied among the nozzle types. Nozzle type did not influence pest (weed, disease and insect) control, or peanut yield (p>0.10) in any of the on-farm trials. These results suggested that peanut growers can utilize these coarser droplet nozzles for pest management in fields with low to average pest pressure during the season. Future research on nozzle evaluation needs to investigate the influence of droplet size, carrier volume, and pressure on coverage and canopy penetration.

Soybean (Glycine max [L.] Merrill) yield losses as a result of plant diseases were estimated by university and government plant pathologists in 29 soybean producing states in the United States and in Ontario, Canada, from 2015 through 2019. In general, the estimated losses that resulted from each of 28 plant diseases or pathogens varied by state or province as well as year. Soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) caused more than twice as much loss as any other disease during the survey period. Seedling diseases (caused by various pathogens), Sclerotinia stem rot (white mold) (caused by Sclerotinia sclerotiorum [Lib.] de Bary), and sudden death syndrome (caused by Fusarium virguliforme O’Donnell & T. Aoki) caused the next greatest yield losses, in descending order. Following SCN, the most damaging diseases in the northern United States and Ontario differed from those in the southern United States. The estimated mean economic loss from all soybean diseases, averaged across the United States and Ontario, Canada was US$45 per acre (US$111 per hectare). The outcome from the current survey will provide pertinent information regarding the important soybean diseases and their overall severity in the soybean crop and help guide future research and Extension efforts on managing soybean diseases.

Aflatoxin production by isolates of Aspergillus flavus varies, ranging from highly toxigenic to completely atoxigenic. Several mechanisms have been identified which regulate aflatoxin production including medium carbon source and oxidative stress. In recent studies, aflatoxin production has been implicated in partially ameliorating oxidative stress in A. flavus. To better understand the role of aflatoxin production in oxidative stress responses, a selection of toxigenic and atoxigenic isolates of A. flavus with moderate to high oxidative stress tolerance were exposed to increasing concentrations of H2O2 in both aflatoxin-conducive and non-conducive media. Mycelial mats were collected for global transcriptome sequencing followed by differential expression, functional prediction, and weighted co-expression analyses. Oxidative stress and medium carbon source had a significant effect on the expression of several secondary metabolite gene clusters including those for aflatoxin, aflatrem, aflavarin, cyclopiazonic acid, and kojic acid. Atoxigenic biological control isolates showed less differential expression under stress than other atoxigenic isolates suggesting expression profiles may be useful in screening. Increasing stress also resulted in regulation of SakA/Hog1 and MpkA MAP kinase signalling pathways pointing to their potential roles in regulating oxidative stress responses. Their expression was also influenced by medium carbon source. These results suggest that aflatoxin production along with that of other mycotoxins may occur as part of a concerted coping mechanism for oxidative stress and its effects in the environment. This mechanism is also regulated by availability of simple sugars and glycolytic compounds for their biosynthesis.

To better understand the evolution of virulence we are interested in identifying the genetic basis of this trait in pathogenic fungi and in developing tools for the rapid characterization of variation in virulence among populations associated with epidemics. Fusarium oxysporum f. sp. vasinfectum (FOV) is a haploid fungus that causes devastating outbreaks of Fusarium wilt of cotton wherever it is grown. In the United States, six nominal races and eleven genotypes of FOV have been characterized based on the translation elongation factor (EF-1α) gene and intergenic spacer region (IGS), but it is unclear how race or genotype based on these regions relates to population structure or virulence. We used genotyping-by-sequencing to identify SNPs and determine genetic diversity and population structure among 86 diverse FOV isolates. Six individuals of Fusarium oxysporum closely related to FOV were genotyped and included in some analyses. Between 193 and 354 SNPs were identified and included in the analyses depending on the pipeline and filtering criteria used. Phylogenetic trees, minimum spanning networks (MSNs), principal components analysis (PCA), and discriminant analysis of principal components (DAPC) demonstrated that races and genotypes of FOV are generally not structured by EF-1α genotype, nor are they monophyletic groups with the exception of race 4 isolates, which are distinct. Furthermore, DAPC identified between 11 and 14 genetically distinct clusters of FOV, whereas only eight EF-1α genotypes were represented among isolates; suggesting that FOV, especially isolates within the widely distributed and common race 1 genotype, is more genetically diverse than currently recognized.

Efforts in genome sequencing in the Aspergillus genus have led to the development of quality reference genomes for several important species including A. nidulans, A. fumigatus, and A. oryzae. However, less progress has been made for A. flavus. As part of the effort of the USDA-ARS Annual Aflatoxin Workshop Fungal Genome Project, the isolate NRRL3357 was sequenced and resulted in a scaffold-level genome released in 2005. Our goal has been biologically driven, focusing on two areas: isolate variation in aflatoxin production and drought stress exacerbating aflatoxin production by A. flavus. Therefore, we developed two reference pseudomolecule genome assemblies derived from chromosome arms for two isolates: AF13, a MAT1-2, highly stress tolerant, and highly aflatoxigenic isolate; and NRRL3357, a MAT1-1, less stress tolerant, and moderate aflatoxin producer in comparison to AF13. Here, we report these two reference-grade assemblies for these isolates through a combination of PacBio long-read sequencing and optical mapping, and coupled them with comparative, functional, and phylogenetic analyses. This analysis resulted in the identification of 153 and 45 unique genes in AF13 and NRRL3357, respectively. We also confirmed the presence of a unique 310 Kb insertion in AF13 containing 60 genes. Analysis of this insertion revealed the presence of a bZIP transcription factor, named atfC, which may contribute to isolate pathogenicity and stress tolerance. Phylogenomic analyses comparing these and other available assemblies also suggest that the species complex of A. flavus is polyphyletic.

Late leaf spot (Cercosporidium personatum) and peanut rust (Puccinia arachidis) are the most important diseases of peanut (Arachis hypogaea L.) in Haiti. Traditional Haitian peanut varieties are not only susceptible to these diseases but are also typically grown without benefit of a fungicide program. Five trials were conducted from 2015 to 2017 to evaluate the performance of six Valencia varieties in Quartier-Morin, Haiti (with an additional trial in 2017 at the Central Plateau) with respect to yield, resistance to rust and leaf spot diseases, and response to a fungicide program. A split-plot design with four or six replications was used in these studies. In each, “variety” was the whole plot and presence or absence of a fungicide program was the subplot. Valencia market types 309 Red, 309 Tan, M2, M3, SGV0801 and a local landrace were compared with and without Muscle ADV (tebuconazole + chlorothalonil, Sipcam) (2.3 L/ha) applied at 45, 60 and 75 days after planting (DAP). Final disease ratings (late leaf spot and peanut rust) were assessed approximately 94 DAP and plots were harvested the day following. In all trials, 309 Tan variety had the least amount of leaf spot and rust, but resulted in the lowest yield in four out of five trials, averaging 1727 kg/ha across fungicide treatments. M3, M2 and 309 Red were generally the numerically highest-yielding varieties, averaging 2906, 2864 and 2541 kg/ha across fungicide treatments, respectively, but were not statistically higher than the local Haitian Valencia, averaging 2374 kg/ha. Three fungicide applications during the season significantly increased yields in most trials for all varieties except 309 Tan. The highest and lowest average increase in yield from fungicide was for 309 Red (1126 kg/ha) and 309 Tan (103 kg/ha), respectively. The results from this study conducted over 2 years and 4 seasons document that while resistance to late leaf spot and rust is available in Valencia varieties, yield potential is not directly associated with that resistance. Also, use of fungicide improves yield potential in more susceptible varieties.

Pre-harvest aflatoxin contamination occurs in maize following kernel colonization by Aspergillus flavus. Aflatoxin contamination resistance is a highly desired trait in maize breeding programs. The identification of novel sources of resistance to pre-harvest aflatoxin contamination is a major focus in germplasm screening efforts. Here, we performed a field evaluation of 64 inbred lines over two years for pre-harvest aflatoxin contamination. Topcrosses were also performed with two testers, B73 and Mo17, to generate 128 F1 hybrids which were also evaluated over two years. Hybrid performance was used to calculate both general combining ability (GCA) of the inbreds, and observed heterosis for aflatoxin resistance. Over both years of the study, aflatoxin concentrations ranged from 80 ± 47 to 17,617 ± 8816 μg kg−1 for inbreds, and from 58 ± 39 to 2771 ± 780 μg kg−1 for hybrids with significant variation between years and lines. The inbred lines CML52, CML69, CML247, GT-603, GEMS-0005, Hi63, Hp301, and M37 W showed

Field, greenhouse, and growth chamber experiments were conducted to determine the level of resistance to Puccinia arachidis Speg. in newly developed breeding lines of peanut (Arachis hypogaea L.). These lines were developed in the UF150 project of the Peanut Collaborative Research and Support Program (Peanut CRSP) as part of the United States Agency for International Development (USAID). Field experiments were carried out in Citra, FL and Tifton, GA from 2010 to 2013. Five genotypes Tifrust-10 and Tifrust-13, and CRSP breeding lines PTBOL3-3, 97x36-HO2-1-B2G-3-1-2-2, and BOL3-7 had the lowest standardized area under the disease progress curve and final disease severity score for rust. The CRSP breeding lines 97x36-HO2-1-B2G-3-1-2-2 and BOL3-7 also appeared to be highly resistant to late leaf spot, caused by Cercosporidium personatum (Berk & M. A. Curtis Deighton). In growth chamber studies, genotypes with longer latent periods generally had lower infection frequencies at 7, 11, and 16 d after inoculation, and smaller percent diseased areas. Latent period and percent diseased area were significantly correlated with stAUDPC. CRSP breeding lines 97x36-HO2-1-B2G-3-1-2-2 and BOL3-7, and plant introductions PI562530, PI568164, and PI298115, were among the genotypes with the lower scores for these components. Several genotypes with multiple disease resistance in different environments and under high disease pressure were identified in these studies. These results indicate sources of rust resistance in the CRSP breeding lines, including several genotypes that could be used as parents in peanut germplasm enhancement programs, and indicate that latent period, percent diseased area, and lesion diameter may be used as indicators for rust resistance in growth chamber studies.

Cotton bacterial blight (CBB), caused by

Frogeye leaf spot (FLS), caused by the fungal pathogen

In peanut (Arachis hypogaea) production, in-furrow applications of the premix combination of the succinate-dehydrogenase-inhibitor (SDHI) fungicide and nematicide fluopyram and the insecticide imidacloprid are used primarily for management of nematode pests and for preventing feeding damage on foliage caused by tobacco thrips (Frankliniella fusca). Fluopyram is also active against many fungal pathogens. However, the effect of in-furrow applications of fluopyram on early leaf spot (Passalora arachidicola) or late leaf spot (Nothopassalora personata) has not been characterized. The purpose of this study was to determine the effects of in-furrow applications of fluopyram + imidacloprid or fluopyram alone on leaf spot epidemics. Field experiments were conducted in Tifton, GA in 2015, 2016, and 2018 to 2020. In all experiments, in-furrow applications of fluopyram + imidacloprid provided extended suppression of early leaf spot and late leaf spot epidemics compared with the nontreated control. In 2020, there was no difference between the effects of fluopyram + imidacloprid and fluopyram alone on leaf spot epidemics. Results indicated that fluopyram could complement early-season leaf spot management programs. Use of in-furrow applications of fluopyram should be considered as an SDHI fungicide application for resistance management purposes.

Cotton bacterial blight (CBB) was a major disease of cotton in the United States in the early part of the 20th century. The recent reemergence of CBB, caused by Xanthomonas citri pv. malvacearum (Xcm) revealed many gaps in our understanding of this important disease. In this study, we employed a field isolate of Xcm from Georgia USA (WT) to generate a non- pathogenic, hrcV mutant lacking a functional Type III Secretion System (T3SS-). We tagged the WT and T3SS- strains with an auto-bioluminescent Tn7 reporter and compared colonization patterns of susceptible and resistant cotton seedlings using macroscopic image analysis and bacterial load enumeration. Wildtype and T3SS- Xcm strains colonized cotton cotyledons of resistant and susceptible cotton cultivars. However, Xcm populations were significantly higher in susceptible seedlings inoculated with the WT strain. Additionally, WT and T3SS- Xcm strains systemically colonized true leaves, although at different rates. Finally, we observed that seed-to-seedling transmission of Xcm may involve systemic spread through the vascular tissue of cotton plants. These findings yield novel insights into potential Xcm reservoirs for CBB outbreaks.

Cotton leafroll dwarf virus (CLRDV) is an emerging virus in cotton production in Georgia and several other Southeastern states in the USA. To better understand the genetic diversity of the virus population, the near complete genome sequences of six isolates from Georgia and one from Alabama were determined. The isolates sequenced were 5,866 nucleotides with seven open reading frames (ORFs). The isolates from Georgia were >94% identical with other isolates from the USA and South America. In the silencing suppressor protein (P0), at amino acid position 72, the isolates from Georgia and Alabama had a valine (V), similar to resistant-breaking ‘atypical’ genotypes in South America, while the Texas isolate had isoleucine (I), similar to the more aggressive ‘typical’ genotypes of CLRDV. At position 120, arginine (R) is unique to Georgia and China isolates, but absent in Alabama, Texas and South American isolates. Ten potential recombinant events were detected in the isolates sequenced. An increased understanding of CLRDV population structure and genetic diversity will help develop management strategies for CLRDV in the USA cotton belt.

Aspergillus flavus and Aspergillus parasiticus produce carcinogenic aflatoxins during crop infection, with extensive variations in production among isolates, ranging from atoxigenic to highly toxigenic. Here, we report draft genome sequences of one A. parasiticus isolate and nine A. flavus isolates from field environments for use in comparative, functional, and phylogenetic studies.

Management of disease affecting peanut in the southeastern United States has benefited from extensive field research identifying disease-associated risk factors since the 1990s. An assessment of risk factors associated with tomato spotted wilt (TSW), caused by tomato spotted wilt virus and spread exclusively by thrips, is available to growers through Peanut Rx, a tool developed to inform peanut management decisions. Peanut Rx provides an assessment of relative TSW risk as an index. The assessment provides information about the relative degree to which a field characterized by a specified suite of practices is at risk of crop loss caused by TSW. Loss results when infection occurs, and infection rates are determined, in part, by factors outside a grower’s control, primarily the abundance of dispersing, viruliferous thrips. In this study, we incorporated meteorological variables useful for predicting thrips dispersal, increasing the robustness of the Peanut Rx framework in relation to variation in the weather. We used data from field experiments and a large grower survey to estimate the relationships between weather and TSW risk mediated by thrips vectors, and developed an addition to Peanut Rx that proved informative and easy to implement. The expected temporal occurrence of major thrips flights, as a function of heat and precipitation, was translated into the existing risk-point system of Peanut Rx. Results from the grower survey further demonstrated the validity of Peanut Rx for guiding growers’ decisions to minimize risk of TSW.

Summary Drought stress conditions decrease maize growth and yield, and aggravate preharvest aflatoxin contamination. While several studies have been performed on mature kernels responding to drought stress, the metabolic profiles of developing kernels are not as well characterized, particularly in germplasm with contrasting resistance to both drought and mycotoxin contamination. Here, following screening for drought tolerance, a drought‐sensitive line, B73, and a drought‐tolerant line, Lo964, were selected and stressed beginning at 14 days after pollination. Developing kernels were sampled 7 and 14 days after drought induction (DAI) from both stressed and irrigated plants. Comparative biochemical and metabolomic analyses profiled 409 differentially accumulated metabolites. Multivariate statistics and pathway analyses showed that drought stress induced an accumulation of simple sugars and polyunsaturated fatty acids and a decrease in amines, polyamines and dipeptides in B73. Conversely, sphingolipid, sterol, phenylpropanoid and dipeptide metabolites accumulated in Lo964 under drought stress. Drought stress also resulted in the greater accumulation of reactive oxygen species (ROS) and aflatoxin in kernels of B73 in comparison with Lo964 implying a correlation in their production. Overall, field drought treatments disordered a cascade of normal metabolic programming during development of maize kernels and subsequently caused oxidative stress. The glutathione and urea cycles along with the metabolism of carbohydrates and lipids for osmoprotection, membrane maintenance and antioxidant protection were central among the drought stress responses observed in developing kernels. These results also provide novel targets to enhance host drought tolerance and disease resistance through the use of biotechnologies such as transgenics and genome editing.

Peanut (Arachis hypogaea) cultivars with resistance or tolerance to Cercospora arachidicola and/or Cercosporidium personatum, the causes of early and late leaf spot, respectively, are needed for organic production in the southeastern U.S. To determine the potential of new breeding lines for use in such production systems, field experiments were conducted in Tifton, GA, in 2014 and 2015 in which nine breeding lines and two cultivars, Georgia-06G and Georgia-12Y, were grown without foliar fungicide applications. In one set of trials, cultivar Georgia-12Y and most of the breeding lines evaluated had early season vigor ratings, early-season canopy width measurements, final plant populations, and pod yield that were greater than those of standard cultivar Georgia-06G. In those trials, final late leaf spot Florida scale ratings were lower and canopy reflectance measured as the normalized difference vegetation index (NDVI), was higher all the breeding lines than those of Georgia-06G. In another set of trials, two of those same breeding lines had final late leaf spot ratings similar to those of Georgia-12Y in 2014, whereas in 2015, six of those breeding lines had final leaf spot ratings that were lower than those of Georgia-12Y. Yields were similar for Georgia-12Y and all the breeding lines in the Gibbs Farm trials. Across years and breeding lines at the Lang Farm, the relationship between visual estimates of defoliation and NDVI was described by a two sector piecewise regression with NDVI decreasing more rapidly with increasing defoliation above approximately 89%. The utility of NDVI for spot comparisons among breeding lines appears to be limited to situations where there are differences in defoliation. Georgia-12Y and multiple breeding lines evaluated show potential for use in situations such as organic production where acceptable fungicides available for seed treatment and leaf spot control are limited.

Annual decreases in soybean (Glycine max L. Merrill) yield caused by diseases were estimated by surveying university-affiliated plant pathologists in 28 soybean-producing states in the United States and in Ontario, Canada, from 2010 through 2014. Estimated yield losses from each disease varied greatly by state or province and year. Over the duration of this survey, soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) was estimated to have caused more than twice as much yield loss than any other disease. Seedling diseases (caused by various pathogens), charcoal rot (caused by Macrophomina phaseolina (Tassi) Goid), and sudden death syndrome (SDS) (caused by Fusarium virguliforme O’Donnell & T. Aoki) caused the next greatest estimated yield losses, in descending order. The estimated mean economic loss due to all soybean diseases, averaged across U.S. states and Ontario from 2010 to 2014, was $60.66 USD per acre. Results from this survey will provide scientists, breeders, governments, and educators with soybean yield-loss estimates to help inform and prioritize research, policy, and educational efforts in soybean pathology and disease management.

Fungicide programs for managing target spot of cotton caused by Corynespora cassiicola were evaluated over 15 site-years in the southeastern United States between 2014 and 2016. Two cultivars, hypothesized to vary in target spot susceptibility, PhytoGen 499WRF (PHY499) and Deltapine 1137B2RF (DPL1137), and four fungicides (azoxystrobin, flutriafol, pyraclostrobin, pyraclostrobin + fluxapyroxad) plus nontreated control, were compared. Fungicide programs consisted of 1) a single application at first flower or disease onset and 2) the first application followed by a second 14 days later. Treatments were applied in a factorial, randomized complete block design. Target spot onset and severity varied among site-years. Except when severity was low, target spot-associated defoliation was greater on PHY499 than on DP1137. Fungicides delayed disease development and defoliation, but application number had little impact. Based on a meta-analysis of 15 site-years, pyraclostrobin-based applications resulted in a 4 to 6% yield preservation, and yield preservation was greater at site-years with early disease onset and >40% target spot associated defoliation. Results suggest a single well-timed application of a pyraclostrobin-based fungicide reduces defoliation and protects cotton yield at locations with high target spot severity. Additional research is needed to identify risk factors for target spot-associated yield losses in cotton production systems.

Previous research has demonstrated the efficacy of prescription fungicide programs, based upon Peanut Rx, to reduce combined effects of early leaf spot (ELS), caused by Passalora arachidicola (Cercospora arachidicola), and late leaf spot (LLS), caused by Nothopassalora personata (syn. Cercosporidium personatum), but the potential of Peanut Rx to predict each disease has never been formally evaluated. From 2010 to 2016, non-fungicide-treated peanut plots in Georgia and Florida were sampled to monitor the development of ELS and LLS. This resulted in 168 cases (unique combinations of Peanut Rx risk factors) with associated total leaf spot risk points ranging from 40 to 100. Defoliation ranged from 13.9 to 100%, and increased significantly with increasing total risk points (conditional R2 = 0.56; P < 0.001). Leaf spot onset (time in days after planting [DAP] when either leaf spot reached 1% lesion incidence), ELS onset, and LLS onset ranged from 29 to 140, 29 to 142, and 50 to 143 DAP, respectively, and decreased significantly with increasing risk points. Standardized AUDPC of ELS was significantly affected by risk points (conditional R2 = 0.53, P < 0.001), but not for LLS. After removing redundant Peanut Rx factors, planting date, rotation, historical leaf spot prevalence, cultivar, and field history were used as fixed effects in mixed effect regression models to evaluate their contribution to leaf spot, ELS or LLS prediction. Results from mixed effects regression confirmed that the selected Peanut Rx risk factors contributed to the variability of at least one measurement of development of combined or separate epidemics of ELS and LLS, but not all factors affected ELS and LLS equally. Historical leaf spot prevalence, a new potential preplant risk factor, was a consistent predictor of the dominant disease(s) observed in the field. Results presented here demonstrate that Peanut Rx is a very effective tool for predicting leaf spot onset regardless of which leaf spot is predominant, but also suggest that associated risk does not reflect the same development for each disease. These data will be useful for refining thresholds for differentiating high, moderate, and low risk fields, and reevaluating the timing of fungicide applications in reduced input programs with respect to disease onset.

Biocontrol using non-aflatoxigenic strains of Aspergillus flavus has the greatest potential to mitigate aflatoxin contamination in agricultural produce. However, factors that influence the efficacy of biocontrol agents in reducing aflatoxin accumulation under field conditions are not well-understood. Shifts in the genetic structure of indigenous soil populations of A. flavus following application of biocontrol products Afla-Guard and AF36 were investigated to determine how these changes can influence the efficacy of biocontrol strains in reducing aflatoxin contamination. Soil samples were collected from maize fields in Alabama, Georgia, and North Carolina in 2012 and 2013 to determine changes in the population genetic structure of A. flavus in the soil following application of the biocontrol strains. A. flavus L was the most dominant species of Aspergillus section Flavi with a frequency ranging from 61 to 100%, followed by Aspergillus parasiticus that had a frequency of 0.05) differences in the frequency of individuals with MAT1-1 and MAT1-2 for clone-corrected MLH data, an indication of a recombining population resulting from sexual reproduction. Population mean mutation rates were not different across temporal and spatial scales indicating that mutation alone is not a driving force in observed multilocus sequence diversity. Clustering based on principal component analysis identified two distinct evolutionary lineages (IB and IC) across all three states. Additionally, patristic distance analysis revealed phylogenetic incongruency among single locus phylogenies which suggests ongoing genetic exchange and recombination. Levels of aflatoxin accumulation were very low except in North Carolina in 2012, where aflatoxin levels were significantly (P < 0.05) lower in grain from treated compared to untreated plots. Phylogenetic analysis showed that Afla-Guard was more effective than AF36 in shifting the indigenous soil populations of A. flavus toward the non-toxigenic or low aflatoxin producing IB lineage. These results suggest that Afla-Guard, which matches the genetic and ecological structure of indigenous soil populations of A. flavus in Alabama, Georgia, and North Carolina, is likely to be more effective in reducing aflatoxin accumulation and will also persist longer in the soil than AF36 in the southeastern United States.

Tomato chlorotic spot virus (TCSV) is emerging as a significant constraint to vegetable and legume crops in the Americas. The complete genome sequence of a TCSV isolate naturally infecting peanut (Arachis hypogea) in Haiti was determined in the effort to build epidemiological knowledge of the virus.

Field trials were conducted in 2015 and 2016 in Tifton, GA to determine the effects of planting dates (24 and 27 April, 4, 11, 19, and 26 May 2015; and 11, 18, and 25 April and 2, 9, and 16 May 2016), peanut (Arachis hypogaea) cultivar (Georgia-06G and Georgia-12Y), and seed treatment (nontreated and treated with azoxystrobin, fludioxonil, and mefenoxam) on epidemics of late leaf spot (Nothopassalora personata), plant populations, and peanut yield. Final severity and AUDPC of late leaf spot increased with later planting dates in both years. For most planting dates in 2015 and the final planting date in 2016, final leaf spot severity and AUDPC were lower for Georgia-12Y than for Georgia-06G. Seed treatment increased plant populations for the 27 April and 4 May planting dates in 2015 and across all other treatments in 2016. Yields were higher for Georgia-12Y than for Georgia-06G in both years. In 2015, yields of both cultivars decreased according to linear functions of day of year of planting date, but there was no effect of planting date on yield in 2016. The combination of early planting with Georgia-12Y shows potential utility for management of leaf spot in situations such as organic production where fungicide use is minimal.

Myrothecium roridum Tode:Fr, pathogenic to a number of cucurbit species, causes fruit rots, cankers on crowns and stems, and leaf spots. Hosts include cantaloupe and honeydew (Cucurbita melo) and cucumber (Cucumis sativus) (1,3). In June 2004, following a period of heavy rainfall, numerous round-to-oblong, brown lesions with concentric rings were observed on leaves of watermelon (Citrullus lanatus) cv. Desert King at the Blackshank Farm in Tifton, GA. Disease was localized in the field and severity was low (5 conidia per ml) plus 0.1% vol/vol Tween 20. Inoculum was applied on leaves and stems until runoff with a hand-held mister, and plants were placed in a dew chamber for 72 h. Ten plants were sprayed with sterile, distilled water to serve as controls. Inoculated and noninoculated control plants were removed from the dew chamber and maintained at 25 to 28°C. Symptoms appeared 8 days after inoculation and were characterized by round, dark lesions with concentric rings; noninoculated plants were symptomless. Sections of symptomatic tissue were plated, and M. roridum was reisolated. Although M. roridum is a common pathogen of melons and cucumber, to our knowledge, this is the first field report of a leaf spot caused by M. roridum on watermelon in the United States. No further occurrences of the disease on watermelon have been observed in Georgia since the initial discovery of M. roridum in 2004; however, losses could be potentially severe if widespread infection of fruit were to occur. References: (1) B. D. Bruton. Crater Rot. Pages 49–50 in: Compendium of Cucurbit Diseases. T. A. Zitter et al., eds. The American Phytopathological Society, St. Paul, MN, 1996. (2) M. B. Ellis. Page 552 in: Dematiaceous Hyphomycetes. CAB International, Wallingford, UK, 1971. (3) D. F. Farr et al. Page 809 in: Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989.

Field experiments were carried out to evaluate the effects of integrated management of early leaf spot, caused by Cercospora arachidicola, and spotted wilt, caused by Tomato spotted wilt virus (TSWV), on peanut (Arachis hypogaea) using host resistance, two tillage systems, and varying fungicide programs. Effects on pod yield and economic return were assessed. Genotypes C-11-2-39 and Tifrunner demonstrated the best field resistance to TSWV, whereas cvs. DP-1 and GA-01R and line C-28-305 were among the genotypes with the best leaf spot resistance. Epidemics of both diseases were comparable or suppressed in strip-tilled plots compared with conventionally tilled plots. Leaf spot intensity decreased with increased fungicide applications, but to a lesser degree with use of resistance and strip tillage. Yields and net returns were similar between tillage treatments in 2002 and lower in strip tillage in 2003. Genotypes with the greatest yields and returns were C-11-2-39, C-99R, and GA-01R. Returns were comparable among the four-, five-, and seven-spray programs in both years, despite differences in yield. The standard production system, Georgia Green in conventional tillage with seven sprays, resulted in lower returns than half the integrated systems tested in 2002, but had comparable or higher returns than nearly all systems in 2003. When significant, yields and returns were correlated with spotted wilt intensity to a greater degree than leaf spot intensity.

Because of the importance of spotted wilt caused by Tomato spotted wilt virus (TSWV), most peanut (Arachis hypogaea L.) breeding programs in the southeastern United States are focusing on developing resistance to TSWV. Many of the cultivars with improved resistance to TSWV are late maturing, requiring 150 days to reach optimum maturity. This factor could greatly impact disease problems at harvest. During November of 2004, an unknown disease was observed on peanut cvs. Georgia 02-C and Hull in a commercial field in Appling County. Symptoms included wilting stems with water-soaked lesions and a dense, gray mold growing on infected tissues. Final disease incidence was less than 5%. For isolation, diseased tissue was surface sterilized by soaking in 0.5% sodium hypochlorite for 1 min, air dried, plated on potato dextrose agar (PDA), and incubated at 20°C. Botrytis cinerea Pers.:Fr., causal agent of Botrytis blight, was isolated from the margins of infected tissue. Mycelia were initially white but became gray after 72 h at which time tall, branched, septate conidiophores formed. Mature, unicellular, ellipsoid, hyaline conidia (8.9 × 10.4 μm) formed in botryose heads (1). Hard, black, irregular-shaped sclerotia formed after 2 weeks. Stems of greenhouse-grown peanut plants (cv. Georgia Green) were inoculated with PDA plugs colonized with either B. cinerea or B. allii Munn. Inoculations were made 3 cm below the last fully expanded leaf on wounded and nonwounded tissue. Noncolonized PDA plugs served as controls (n = 9). Plants were arranged in a dew chamber at 20°C in a randomized complete block design. Lesions and spore masses identical to those observed in the field appeared 3 to 5 days after being inoculated with B. cinerea. The B. allii inoculations caused only superficial lesions. After 5 days, mean lesion lengths for B. cinerea were 59 and 37 mm for wounded and nonwounded inoculations, respectively. B. cinerea was recovered from 100% of the symptomatic tissues. Botrytis blight is considered a late-season disease that occurs in cool, wet weather (3). Symptoms similar to those of Botrytis blight were observed on mature and over-mature peanut in Georgia and have been cited as “unpublished observations” (2); however, to our knowledge, this is the first report of the disease in Georgia. Although Botrytis blight is not considered a major peanut disease, it may become more prevalent at harvest as producers utilize late-maturing cultivars to manage spotted wilt. References: (1) H. L. Barnett and B. B. Hunter. Illustrated Guide of Imperfect Fungi. 4th ed. The American Phytopathological Society, St. Paul, MN, 1998. (2) K. H. Garren and C. Wilson. Peanut Diseases. Pages 262–333 in: The Peanut, the Unpredictable Legume. The National Fertilizer Assoc. Washington D.C. 1951. (3) D. M. Porter. Botrytis blight. Pages 10–11 in: Compendium of Peanut Diseases. 2nd ed. N. Kokalis-Burelle et al., eds. The American Phytopathological Society, St. Paul, MN. 1997.

Southern rust, caused by Puccinia polysora Underw., occurs frequently on corn (Zea mays) grown in subtropical or tropical regions. When conditions are favorable, southern rust also occurs in temperate climates of the central and southern United States although the fungus does not survive on corn crop residue and must be introduced to temperate regions each growing season. Several single, dominant, resistance genes, designated as Rpp genes, convey hypersensitive, chlorotic fleck reactions when challenged with avirulent isolates of P. polysora (1). Rpp resistance prevents or limits the formation of uredinia. The Rpp9 gene has been used successfully in North America in the past 20 years to control southern rust even though the gene has been ineffective in other parts of the world (e.g., Africa and Hawaii) because of the prevalence of virulent races. During the past 3 years, Rpp9 virulence has occurred in the western hemisphere (e.g., Brazil, Mexico, Nebraska, and Texas), but prior to 2008, uredinia were not observed east of the Mississippi River on corn with the Rpp9 gene. A few uredinia were observed on corn with the Rpp9 gene in eastern Nebraska in 2006 and near Victoria, TX in 2007 (W. Dolezal, personal observation). In July of 2008, a virulent isolate of P. polysora was confirmed from Grady County, GA on corn lines with the Rpp9 gene including the original source of this resistance gene, Boesman yellow flint, which is PI 186208 (3). In August of 2008, isolates of P. polysora were collected from severely infected corn hybrids with Rpp9 grown in Macon County, GA. Rust samples from hybrids without Rpp genes also were collected in Burke County, GA where Rpp-resistant corn was asymptomatic. In greenhouse trials, corn lines with and without the Rpp9 gene were inoculated with urediniospores from collections from Burke and Macon counties and Illinois. Rust infection types (1) were scored 18 to 25 days after inoculation. The Macon County isolate produced type 1 and 2 infections (small uredinia surrounded by necrotic or chlorotic tissue) on Oh43Rpp9 and W64aRpp9 and type 4 infections (large, sporulating uredinia) on two versions of a commercial hybrid with and without the Rpp9 gene and on Va59 (which carries an Rpp gene different from Rpp9). The Burke County isolate and an isolate from Illinois collected in 2001 produced type 0 infections (chlorotic flecks) on all of these lines except the non-Rpp version of the commercial hybrid which had a type 4 reaction. To our knowledge, Rpp9-virulent isolates of P. polysora have not been reported from the continental United States for nearly 50 years. In the late 1950s and early 1960s, A. L. Robert (2) collected isolates of P. polysora from throughout the world and observed multiple races on a set of host differentials that is no longer available. A. L. Robert's collection included an isolate from Georgia that was virulent on PI 186208. Commercial hybrids containing the Rpp9 gene may continue to be resistant throughout most of North America if previously common Rpp9-avirulent isolates of P. polysora are prevalent, but those hybrids should be carefully monitored for infection by newly introduced Rpp9-virulent isolates. References: (1) A. L. Hooker. Page 207 in: The Cereal Rusts. Vol. II. Academic Press, San Diego, 1985. (2) A. L. Robert. Phytopathology 52:1010, 1962. (3) A. J. Ullstrup. Phytopathology 55:425, 1965.

Field experiments were conducted at Gainesville and Marianna, FL in 2004 and 2005 in which severity of spotted wilt, caused by Tomato spotted wilt virus, and pod yield were compared in six peanut (Arachis hypogaea) cultivars. The six cultivars included the moderately field resistant cultivars ANorden, C-99R, and Georgia Green; the highly field resistant cultivars AP-3 and DP-1; and the susceptible cultivar SunOleic 97R. There were four trials at each location, with four planting dates that ranged from late March to early June. Tomato spotted wilt severity in moderately resistant and susceptible cultivars was lower at Gainesville than at Marianna in both years in moderately resistant and susceptible cultivars. Trends in incidence for the two locations were less evident for AP-3 and DP-1. At Gainesville, there were few differences in tomato spotted wilt severity, and severity ratings were similar for Georgia Green and SunOleic 97R in two of four trials in 2004 and across all trials in 2005. At Marianna, severity ratings were lower for Georgia Green than for SunOleic 97R in six of the eight trials, and severity of tomato spotted wilt was lower for AP-3, C-99R, and DP-1 than for Georgia Green in all eight trials. In 2004, there was a trend toward decreasing severity ratings for Georgia Green and SunOleic 97R with later planting dates, but not for AP-3 or DP-1 at Marianna. Split-plot field experiments were also conducted at Tifton, GA in 2005 through 2007 in which incidence of tomato spotted wilt and pod yield were compared for peanut cultivars AP-3 and Georgia Green across planting dates ranging from late April through late May. Incidence of tomato spotted wilt was lower for AP-3 than for Georgia Green within each planting date of all years, and planting date effects were smaller in AP-3, if observed at all, than in Georgia Green. In most planting dates of all three trials, yields were higher for AP-3 than for Georgia Green. The relationships between yield and planting date were not consistent. These results indicate that the level of field resistance in AP-3 and DP-1 cultivars is sufficient to allow planting in late April without greatly increasing the risk of losses to tomato spotted wilt.